The invention relates to natural pharmaceutical chemistry, especially relates to kernel oils extracted from plant kernels, more especially relates to kernel oils extracted from spine date (Semen ziziphi spinosae), flatspine pricklyash and walnut (Semen persicae), the process for preparing same, the pharmaceutical composition containing same and the medicinal use thereof.
The spine date kernel and the flatspine pricklyash are Chinese traditional medicines for both medical- and food-purpose, which have functions such as nourishing liver, calming heart, constraining sweating, engendering liquid, warming the middle warmer to alleviate pain, killing parasites and relieving itch.
The walnut kernel is one of the Chinese traditional medicines for both medical- and food-purpose, and has functions of nourishing the kidney, warming the lung and moistening the bowel. However, walnut kernel oil has usually been used as an edible oil and has not been reported as a parenteral nutritional agent. The common walnut kernel oil preparation is not advantageous to human absorption, and its medical application is restricted since it is difficult for it to meet the clinic requirements as an intravenous emulsion. The backward technology of extraction and separation in the prior art induces in unqualified chemical and physical properties (such as acid value) and high impurity contents.
In the prior art, the fat emulsions prepared from the soya bean oil, the cotton seed oil and the red flower oil are commonly used to replenish nutrients and energy to human body. The fat emulsion made from soya bean oil is more widely used. However, no work on the improvement of immunological functions, the increase of serum proteins, the inhibition on transplanted tumor, experimental Lewis lung cancer and liver cancer HAC, and the functions of nourishing kidney, warming lung and loosening bowel has not been reported in the studies of the fat emulsion made from the soya bean oil.
Thus, the first aspect of the invention is to provide a kernel oil from kernels of plants which can be used for the preparation of a novel intravenous emulsion having functions of replenishing nutrients, improving immunologic function and increasing serum proteins.
The second aspect of the invention is to provide two special technological processes for the extraction of the kernel oil of the invention.
The third aspect of the invention is to provide a pharmaceutical composition containing the kernel oil of the invention, including intravenous emulsion and oral capsules. Said intravenous emulsion can replenish nutrients, improve immunologic function and increase serum proteins. Also, it has functions of nourishing kidney, warming lung and loosening bowel. In addition, it is not expensive.
The fourth aspect of the invention is to provide the use of the kernel oil of the invention in the preparation of remedies for the treatment of diseases such as tumors, AIDS and hypoimmunity, etc.
The kernel oil extracted from plant kernels according to the first aspect of the invention comprises triglycerides 90-99.9%, diglycerides 0.01-5%, monoglycerides 0.01-3%, sitosterol 0.1-2.5% and cyclolanosterol 0.01-1%.
One process for extracting the kernel oil of according to the invention comprises the steps of:
1) Crude extraction: the kernel/kernel powder being expressed or extracted with an organic solvent or via supercritical fluid extraction to obtain a crude oil;
2) Decoloring: decoloring the crude oil with an adsorbent decoloring agent to obtain a decolored oil;
3) Caustic refining: dissolving the decolored oil in petroleum ether, adding a stoichiometric amount of NaOH under agitation, standing still and demixing, then washing the organic phase, thus obtaining an emulsion;
4) Demulsifying: adding acetone into the emulsion with agitation, separating layers and achieving the upper layer of oil phase.
5) Adsorption and water-washing: Subjecting the oil phase to be adsorbed with neutral alumina and kaolin sequently, and filtered, then removing the organic solvent from the filtrate in nitrogen atmosphere, and washing the oil phase with worm water, drying, then adsorbing with neutral alumina, thus obtaining a refined oil.
Another process for extracting the kernel oil according to the invention comprises the steps of:
1) Crude extraction: the kernel/kernel powder being expressed or extracted with an organic solvent or via supercritical fluid extraction to obtain a crude oil;
2) Degumming: agitating and heating the crude oil, then adding phosphoric acid to render a complete reaction;
3) Caustic refining: adding NaOH or Na2CO3 solution at the same temperature into the degummed oil to render a complete reaction, then standing still and demixing, thus obtaining a refined oil;
4) Water-washing: washing the caustic-refined oil with pure water to obtain a water-washed oil.
5) Dewatering: adding an adsorbent into the water-washed oil or heating the water-washed oil under vacuum to remove the water, thus obtaining a clear dewatered oil;
6) Decoloring: decoloring the dewatered oil with an adsorbent decoloring agent to obtain a decolored oil;
7) Deodorizing: heating the decolored oil with agitation under vacuum in nitrogen atmosphere to raise the oil temperature to 120-160xc2x0 C., feeding steam made from pure water into the oil and further heating the oil to 160-260xc2x0 C. and keeping for 0.5-2 hours, then cutting short the pure water steam, thus removing the moisture from the oil and obtaining deodorized oil.
2. The pharmaceutical composition according to the invention comprises a therapeutically effective amount of the kernel oil extracted from kernels of plants and one or more pharmaceutically acceptable adjuvants.
The invention also provides the use of the kernel oil from kernels of plants in the preparation of remedies for treating tumors, AIDS, hypoimmunity, infantile malnutrition, postoperation and diseases in need of supplementing fat elements.
The kernel oil extracted from plant kernels according to the invention is a clear light yellow oily liquid which comprises triglycerides 90-99.9%, diglycerides 0.01-5%, monoglycerides 0.01-3%, sitosterol 0.1-2.5% and cyclolanosterol 0.01-1%.
The lipolysis of said kernel oil gives the following fatty acids: hexadecanoic acid 5-8%, octadecanoic acid 1-3%, octadecenic acid 18-30%, octadecadienoic acid 50-65% and calendic acid 6-14%.
The kernel oil of the invention possesses the following physical properties examined on the basis of fatty oil: relative density 0.920-0.930, refractive index 1.470-1.480, acid value less than 0.80, iodine value 120.0-155.0, saponification value 180.0-200.0, peroxide value less than 30.0 meq.kgxe2x88x921, ignited residue 0.01-0.04%, arsenic salts less than 2 ppm, heavy metals less than 10 ppm, and mean molecular weight 873.96.
The preferred embodiments of the kernel oils of the invention are those extracted from kernels of spine date, flatspine pricklyash and walnut.
With the walnut kernel as an example, the first process for extracting kernel oils includes the following steps:
Primary extractionxe2x80x94The kernel/kernel powder being expressed or extracted with an organic solvent or via supercritical fluid extraction to obtain a crude walnut kernel oil;
Decoloringxe2x80x94Adding proper amount of petroleum ether into the crude oil and thoroughly mixing it, adding proper amount of activated carbon for injection-purpose, keeping the temperature constant, filtering the mixture, and recovering the petroleum ether, thus obtaining a decolored oil;
Caustic refiningxe2x80x94Putting the decolored oil and a proper amount of petroleum ether into a reaction kettle, adding 2% NaOH in an amount calculated on basis of the acid value and the amount of the decolored oil with agitation, standing still, and removing the lower layer of liquid waste, then adding two fold of warm distilled water with agitation, standing still, and removing the lower layer of liquid waste, then washing once more in the same method, duration and water temperature as above, sufficiently standing still, removing the lower layer of liquid waste, and obtaining the upper layer of emulsion;
Demulsifyingxe2x80x94Metering and transferring the emulsion layer into a separator, and filling acetone in a given ratio to the amount of emulsion layer with constant agitation, after standing still and separating layers, removing the lower layer of liquid waste, thus obtaining the upper layer of oil phase.
Water-washingxe2x80x94Adding the neutral heat-activated alumina in an amount based on the oil amount to conduct the adsorption, mixing thoroughly, standing still and filtering to obtain a clear oil, then preheating the metered clear oil in a reaction kettle, adding necessary amount of the heat-activated white bole (kaolin) with agitation and heat preservation, filtering under vacuum, heating the filtrate in a washing vessel, removing the organic solvent in nitrogen atmosphere, filling warm distilled water in an amount base on the oil amount, mixing thoroughly, standing still, and removing the lower layer of liquid waste, then heating the oil layer in nitrogen atmosphere to dewater and dry it until the oil layer becomes clear, then adding neutral activated alumina in an adequate amount, mixing thoroughly, standing still, and making sterile filtration, thus obtaining a refined oil.
The purpose of protection with nitrogen in the above process is mainly for improving oil quality through preventing the fat from oxidation and controlling fat""s peroxide value.
If desired, the kernel oil from walnut obtained above can further be packed and sterilized. Then it can be filled into specified containers and sterilized, thus a kernel oil for injection is obtained.
The second process for extracting the kernel oil of the invention includes the following steps:
1) Crude extraction: the kernel/kernel powder being expressed, or extracted via supercritical fluid extraction to obtain a clear crude oil;
2) Degumming: Putting the crude oil into a reaction kettle, filling nitrogen, agitating, heating, adding a proper amount of phosphoric acid, and agitating fast to render a complete reaction;
3) Caustic refining: Directly adding NaOH or Na2CO3 solution with the same temperature into the degummed oil mixture in nitrogen atmosphere, agitating fast to enable free fatty acids to react completely, then standing still to separate layers in the condition of heating and filling nitrogen, and removing the soapstock to obtain a refined oil.
4) Water-washing: Adding proper amount of sodium chloride solution into the caustic-refined oil under agitating, standing still to separate layers with the keeping of temperature, and removing the lower layer of liquid waste, then washing twice with pure water in the same method, duration and water temperature as above, standing still to separate layers and removing the lower layer of liquid waste to obtain a water-washed oil;
5) Dewatering: Adding proper amount of activated alumina into the water-washed oil, mixing thoroughly, standing still and filtering the mixture to obtain a clear dewatered oil;
6) Decoloring: Adding the dewatered oil into a stainless steel reaction kettle, mixing and heating the oil up to 140xc2x0 C. with agitation under vacuum in nitrogen atmosphere, adding proper amount of a mixture of activated carbon and activated kaolin, agitating thoroughly under vacuum at 80-90xc2x0 C., cooling, and filtering the mixture thus obtaining a decolored oil.
7) Deodorizing: Adding the decolored oil into a stainless steel reaction kettle, heating the decolored oil up to 140xc2x0 C. with agitation under vacuum in nitrogen atmosphere, then feeding steam made from pure water in stead of nitrogen, and further heating the oil up to 190xc2x0 C. and keeping the same temperature for 1.5 hours, then cutting short the pure water steam, and agitating in nitrogen atmosphere again, then cooling the oil to obtain a deodorized oil.
If desired, the obtained deodorized oil can further be sterilized, i.e., the deodorized oil is introduced into a stainless steel sterilizing vessel and heated up to 160xc2x0 C. with agitation under vacuum. After keeping at this temperature for 2 hours, the oil is cooled down and aseptically filtrated, then filled into containers in nitrogen atmosphere and sealed, thus obtaining a kernel oil for injection.
The pharmaceutical composition according to the invention comprises a therapeutically effective amount of the kernel oil of the invention extracted as above and one or more pharmaceutically acceptable adjuvants.
The pharmaceutically acceptable adjuvants in the composition of the invention include one or more substances selected from a group consists of an emulsifier, a solubiliser, a latent solvent, an isotonic regulator, an antioxidant and an stabilizer.
The pharmaceutical composition of the invention can further comprise other active drugs selected from anti-tumor agents, anti-AIDS agents, immunomodulators and nutrients.
The pharmaceutical composition of the invention can further comprise one or more vegetable oils, e.g., copra oil, peanut oil, etc.
The pharmaceutical composition of the invention can be a fatty emulsion, such as an intravenous fat emulsion or an oral fat emulsion. It can also be a soft capsule or an iodine oil, etc.
One of the preferred embodiments of the pharmaceutical composition of the invention is a fatty emulsion, wherein said pharmaceutically acceptable adjuvants are an emulsifier and an isotonic regulator. More preferred embodiment is an intravenous fatty emulsion of walnut kernel oil.
Preferably, the intravenous fatty emulsion of walnut kernel oil contains, based on the total volume of the emulsion of 100 ml, from 5 g to 30 g, more preferably from 10 g to 30 g, most preferably 20 g of parenteral walnut kernel oil (walnut kernel oil for injection).
The emulsifier used in this invention can be phosphatide (including soya lecithin, yolk lecithin and soya phosphatide), Pluronic, polyglyceryl dipalmitate, etc. It is preferable to use natural emulsifiers, e.g., soya lecithin or yolk lecithin, in the intravenous fatty emulsion in an amount, based on the total volume of the emulsion of 100 ml, of bout 1.0-3.0 g, more preferable, 1.0-2.0 g, and most preferably, 1.2 g.
The isotonic agent used in the invention can be glycerin, sorbitol, xylitol and glucose, etc., preferably, glycerin in an amount, based on the total volume of the emulsion of 100 ml, from 1.5 g to 3.0 g, more preferably, 2.5 g.
The invention is further described with examples, which are presented for purpose of illustration and by no way of limitation to the invention.